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PubMed1971

Flow dichroic spectra of tobacco mosaic virus and their protein assemblies.

Ultraviolet flow dichroisms of tobacco mosaic virus (TMV), TMV-RNA and TMV-protein were measured using three strains of TMV. 1. Large positive dichroisms were observed with three strains of TMV, namely ordinary, bean form of bean, and tomato strains (TMV-OM, TMV-B and TMV-T, respectively) at about 255, 276, 284 and 290 nm. The positive dichroisms were confirmed with reconstituted protein assemblies of TMV-OM and TMV-B at about 276, 284 and 290 nm where tyrosine and tryptophan residues of these proteins contribute. These results show that the electronic transition moments of their base groups and aromatic groups are nearly parallel to the polymer axis. It is suggested that there is a strong interaction between the base groups of RNA and aromatic groups of amino acid residues in TMV. 2. A small negative dichroism was observed near 296-300 nm with intact TMV-OM and TMV-T and with the reconstituted protein polymer of TMV-OM. But negative dichroism was not observed either with the intact virus or the protein assembly of TMV-B. 3. Isolated RNA from TMV-OM, TMV-B and TMV-T showed no dichroism. The configuration of RNA in TMV appears to be imposed on it by its packing with the protein.

Circular Dichroism↗
PubMed1993

Effects of the tom1 mutation of Arabidopsis thaliana on the multiplication of tobacco mosaic virus RNA in protoplasts.

For the multiplication of RNA viruses, specific host factors are considered essential, but as of yet little is known about this aspect of virus multiplication. To identify such host factors, we previously isolated PD114, a mutant of Arabidopsis thaliana, in which the accumulation of the coat protein of tobacco mosaic virus (TMV) in uninoculated leaves of an infected plant was reduced to low levels. The causal mutation, designated tom1, was single, nuclear, and recessive. Here, we demonstrate that the tom1 mutation affects the amplification of TMV-related RNAs in a single cell. When protoplasts were inoculated with TMV RNA by electroporation, the percentage of TMV-positive protoplasts (detected by indirect immunofluorescence staining with anti-TMV antibodies) was lower (about 1/5 to 1/10) among PD114 protoplasts than among wild-type protoplasts. In TMV-positive PD114 protoplasts, the amounts of the positive-strand RNAs (the genomic RNA and subgenomic mRNAs) and coat protein reached levels similar to, or slightly lower than, those reached in TMV-positive wild-type protoplasts, but the accumulation of the positive-strand RNAs and coat protein occurred more slowly than with the wild-type protoplasts. The parallel decrease in the amounts of the coat protein and its mRNA suggests that the coat protein is translated from its mRNA with normal efficiency. These observations support the idea that the TOM1 gene encodes a host factor necessary for the efficient amplification of TMV RNA in an infected cell. Furthermore, we show that TMV multiplication in PD114 protoplasts is severely affected by the coinoculation of cucumber mosaic virus (CMV) RNA. When PD114 protoplasts were inoculated with a mixture of TMV and CMV RNAs by electroporation, the accumulation of TMV-related molecules was approximately one-fifth of that in PD114 protoplasts inoculated with TMV RNA alone. No such reduction in the accumulation of TMV-related molecules was observed when wild-type protoplasts were inoculated with a mixture of TMV and CMV RNAs or when wild-type and PD114 protoplasts were inoculated with a mixture of TMV and turnip crinkle virus RNAs. These observations are compatible with a hypothetical model in which a gene(s) that is distinct from the TOM1 gene is involved in both TMV and CMV multiplication.

Arabidopsis↗